Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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USE OF 1,2,4-TRIAZOLE DERIVATIVES FOR THE MANUFACTURE OF A MEDICAMENT FOR THE
TREATMENT OF CANCERS
TECHNICAL FIELD
s This invention is a ph~.. A~c~ co,--posilion that inhibits the growth of
cancers, le~emi~ and tumors in mAmmAIc particularly in human and warm blooded
Anim-Alc The co---posilion co~ c a lH-1,2,4-triazole derivative. The co...pos;lions
can also be used to treat viral inf~ctiQnc
BACKGROUND OF THE INVENTION
Cancers are the leading cause of death in animals and hllm~nc The exact cause
of cancer is not known, but links ~ ,.. certain activities such as cmol i~ or
C-I~GS.J~ to carri~ogpnc and the ;~ id~ ~ce of certain types of cancers and tumors has
been shown by a number of ~-,se~
Many types of c~ ,ulic agents have been shown to be ~
5 against cancers and tumor cells, but not all types of cancers and tumors r~_s~,vl d to
these agents. U ~- lu~dlely, many of these agents also destroy normal cells. Theexact .~eel-An ~ for the action ofthese ~ olhe~apeutic agents are not always
'~u~own.
Despite advances in the field of cancer Ir~ e.~l the leading ~ s to date
20 aresurgery"~ ;o~andcl- o~ .a~,~. Ch ~~011~ d~ UI;Capple- ~9 are_aidto
fight cancers that are . ~ d or ones that are p&~ Lul~ a~ . Such
~,~,locidsl or ~ o~hc agents work best on cancers with large growth factors, i.e.,
ones whose cells are rapidly dividing. To date, ho....ones, in particular eSllv3_.l,
pn,gc~t~rone and t~ slv,i~ -vne, and some ' ~ produced by a variety of
2s ubes, aLk~l~;nl3 agents, and anti-mPt~holitps form the bulk of 11.~ s available to
Ideally ~ lolo,ic agents that have ~l e ;l~ y for cancer and tumor cells
while not ~ normal cells would be e~ d~ le U ~. Iundtely, none
have been found and instead agents which target e ~p~ rapidly dividing ceUs (both
tumor and normal) have been used.
Clearly, the d~ O~ ~ ~1 of materials that would target tumor ceUs due to
some unique s~ y for them would be a breakll--ougl-. Alternatively, materials
that were ~OIGA;C to tumor ceUs while eA~,. I.-~g mild effects on normal ceUs would be
des.. bl~ Th~. ~r~. e, it is an object of this invention to provide a ph& ~ ti
co...po~;l;on that is e ~cl;~._ in i~-k ~. I;.~g the growth oftumors and cancers in
3s ' with mild or no effects on normal ceUs.
More ~l e~ , it is an object of this invention to provide an anti-cancer
o - ~;o~ co...l..;r -~g a ~ c~Jti~Al carrier and a lH-1,2,4-tri~le derivative as
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defined herein along with a method for treating such cancers
These and other objects will become evident from the following detailed
desc. ;~tiOII of this inv~ .-Lions.
SI~ARY OF THE rNVENI ION
Aph~.. ~c~tic~lco.--l,o,;~ionforL.~ ofm~mm~lc andinparticular,
warm blooded animals and hum~nc co--.~ .is;n~ a pha....~c~ltic~l carrier and an
effective amount anti-cancer compound sele. led from the group co~ g of:
~/z\
O O
C --C /
~h.,~ Z is an alkylene s~le~led from the group coric ~ of
CH2-CH2-,-CH2-CH2-CH2-, -CH(CH3)-CH(CH3)- and -CH2-CH(alkyl) v~L~r~
said alkyl has from 1 to about 10 carbon atoms; and Ar is a ... ..h~r s~lP~lcd from the
group cc~nc ~ 3 of phenyl, svb ~I;lvlcd phenyl, thienyl, h~ 1, naphthyl and
nuo~ .h~,.e..l "~vl~sl;~ ed phenyl" has the ~ 3n ~g of a phenyl radical having
thereonfrom 1 to 3 s~ s~ e~ indeponf~ntlyfromthe group ÇQ~ of
5 halo, lower alkyl, lower alkyloxy, cyano and nitro. The Ll,e.~ lti~lly active acid
~d~iti~n salts ofthe rO.~,goi~u co~..po ~ ~d (I) are also embraced within the scope ofthis
Li~
As used in the fc,l~3Oul~ d~ ;on of Z, the term "alkyl" is meant to include
straiBlt and branch chained l.~dl~)ca.l,on radicals having from l to ahout l0 carbon
20 atoms, such as, for ~ . 'e, methyl, ethyl, 1-l..~ Lhjl.,LI.~I, propyl, 1,l-du.._Ll.~lethyl,
buty1, pentyl, hexyL heptyl, octyl, decyl and the lilce; as used herein "lower alkyl" may
be ~raight or branch chained sdLu.dted hydrocarbons having from l to 6 carbon atoms,
such as, for; , ' e, methyl, ethyl, propyl, 1 -methylethyl, butyl, 1, l-diu~ll.;lcll.~rl,
pentyl, he~cyl and the ILce alkyls; and the term "halo" is generic to h~log~n atoms of
2S atomic weight les_ than 127; i.e., fluoro, chloro, bromo and iodo.
These cc . ~ innQ can be used to inhibit the growth of cancers and other
tumors in humans or animals by z ~ Lon of an effective amount either orally,rectally, t~r ~ y or pa.~.lt .~.lly, intravenously or by i jection into the tumor. These
cc,..~l os l;~nQ do not c;~ ~ ,lly affect healthy cells a_ cc,..lpared to adl;a...~cin which
30 has a d~,~lull_nlal effect on healthy cells
These CC~--.pG .;Lol1s are also be used to treat vinuses
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DETAlLED DESCRIPTION OF THE INVENTION
A. Dsr jti~ -
As used herein, the term "co.l-~-is;ng" means various co.npon_..ls can be
conjointly employed in the phal .~we. tic,ql co---~,osilion of this invention. Accordingly,
s the terms "conc:cl;~.g eCsenti-q-lly o~' and "CQI~C ~ .g of' are embodied in the term
~. . .
Cf~
A~s used herein, a "~hdweutiç-q-lly acccpt~le" coror ...................................................................l is one that is
suitable for use with humans and/or animals without undue adverse side effects (such
as to~cily, irritation, and allergic ,.,~,onse) co.. --~n~ ate with a rcas~--'lebenefit/risk ratio.
As used herein, the term "safe and effective amount" refers to the q~ lily of a
co...ron-,-.l which is snffici~nt to yield a desired th_~ay_ulic 1~ ~pon~ without undue
adverse side effects (such as toxicity, h.ilaliOI~ or allergic ~3p~ C)CO~ t~
with a reas~r?b'e benefit/risk ratio when used in the manner ofthis inve..lio.l. The
5 specific "safe and effective ~no~ will, obviously, va~y with such factors as the
particular con-l;l;on being treated, the ph~;,;cal con~);l;~n ofthe patient, the type of
"'-" ""21 being treated, the duration of the 11 ~ - ~ n the nature of cor.~;u-,~ therapy
~i~any~ and t~e specific ~....-1~ ,.c emptoyed and the structure ofthe comro~nfl~ or
its derivatives.
As used herein, a "phal .--w~ a~ itisn salts" is salt of the anti-cancer
c~- -~l-ol~-.fl with an organic or ino~ ~ c wid. These pl~f~ d acid r d~ isn salts are
c'-1~ idcs, bromides, ~ t.~<;, nitrates, ph~ k'~$, s~lr~ t~, fol '-g, IL.I.~.tes,
f~ , mqlqte~, citrates, bcn~o-~c~, sali~lahs, ascoll,alcs, and the lilce.
As used hercin, a "ph~ ~ ql carrier" is a ph~.-.~ lly a~ c IFt~b'e
2s solvcn~ f~ R agent or vehicle for delivering the anti-cancer agent to the animal
or hun~rL The carrier may be liquid or solid and is s~leeled with the planned manner
of admi~ lion in mind.
As used herein, ~cancer" refers to all types of cancers or ncoplasll. or m~igr ~tumors and all types of cancers inrl~lfling le~ m;q that are found in mqmm~t-
Y~) As used herein, the "anti-cancer c ~j--.pOi~ lcn are the lH-1,2,4-tli; 1~ and
their salts. The exact 1H-1,2,4-~ s are de~_.;l.ed in detail below. The p~.-~
~lldte-;&ls are the plodu~,ls sold under the names ~plOp ~ ~" by Janssen
Ph& ~C NV (n,.l~;.. ),
As used herein, "viruses" inrl~des viruses which cause ~ s (viral
inff~ctif ne) in man and other warm bl~oded ~nimqle, such as HlV virus, herpes,
i..n.,,.., and rhinoviruses.
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B. T~E ANTI-CANCER COMPOUNDS
The anti-cancer compounds are lH-1,2,4-triazole derivatives which are known
for their Antifimg~l activities. They are systemic materials used to prevent anderadicate fungi. The c~ o~ c have the following structure:
~ /Z\
0~ ~0
CH2--C Ar
wL_rd.~ Z is an alkylene ~ ~ eil from the group coh~ ling of
CH2-CH2-,-CH2-CH2-CH2-, -CH(CH3)-CH(CH3)- and -CH2-CH(alkyl) wherein
said alkyl has from I to about 10 carbon atoms; and Ar is a ~ ,...b. . sPle~l~d from the
group con~ R of phenyl, ~-~I,.t;l.lied phenyl, thienyl, hAlot~ yl, ..aphll-~l and
0 fluorenyl, .. h_n,.n ".~Jb,~ Qd phenyl" has the ~ ~An ~g of a phenyl radical having
thereon from 1 to 3 ~ ~l~le~ lcp~ ~d~ nl~r fromthegroup co~ of
halo, lower alkyl, lower alkyloxy, cyano and nitro. The IL_.~p~ active acid
A~ ion salts ofthe fong; ~ co-..l.ù~ (1) are also e...~. ced within the wope ofthis
i,.~e.,Lon.
As used in the fole~.oin~. d~ ;o~- of Z, the term "alkyl" is meant to include
straight and branch chained L~dlù.,a,l,on radicals having from 1 to about 10 carbon
atoms, such as, for . p'e, methyl, ethyl, l-methylethyl, propyl, l,1-d",._ll.~lethyl,
butyl, pentyl, hexyl, heptyl, octyl, decyl and the like; as used herein "lower alkyl" may
be straight or branch chained al~i l.~d,oc~l,ons having from 1 to 6 carbon atoms,
20 such as, for ~ . 'e, methyl, ethyl, propyl, l-methylethyl, butyl, 1, l-dil~ ,LI.~I,
pentyl, hexyl and the lilce alkyls; and the term "halo" is generic to h~log~n atoms of
atomic wdght less than 127; i.e., fluoro, chloro, bromo and iodo.
Their pl~ ~ lly ac e -r ~bl e acid a'~itinn salts with both organic and i~lurL '-
acit~ can also be used herein.
2s ~f~ d d~;~t;._s include:
1-[2-(2,4~:c''- ùph_..~rl)-1,3~ n-2-yl"-_~ l]-lH-1,2,4-l, 1~,
l-t2-(2,4-.' '', ophc.. ~l)~methyl-1,3-~ 2-~ l]-lH-1,2,4-i - ~le
l-t2-(2,4- " ' '- u?l~ l 1,3~ -2-yl~ lL~1]-1H-1,2,4-l~ :'e
1-[2-(2,4-d ' '~ upl-~ 1)-4-propyl-1,3-~i, ol~-2-ylmethyl]-lH-1~2~4-t ~le,
1-[2-(2,4~ ' lo~pl~ l)-4-pentyl-l~3-~ ynl~n-2-~ l]-lH-l~2~4-ll ~1~ and
the ~ ti~lly active acid ~d~litior~ salts thereo~
These co---l-o~ s are p~ ,d acco,d., ~ to the method desc.il~ed in U.S.
4,079,062 issued to Van Reet. et al, Mar 14, 1978.
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s
It is believed that these particular materials have the capability of red~lc;~lgtumors or de~".;~ing their growth signifiç~ntly bec~ql~ee oftheir ability to inhibit the
synthesis of sterols.
C. DOSAGE
Any suitable dosage may be given in the method of the invention. The type of
disease (cancer, lell~miq or virus), the co---po~ d, the carrier and the amount will
vary widely dep~ n~ g on the species of the warm blooded animal or human, body
weight, and tumor being treated. Generally a dosage of ~ about 2 milligrams
(mg) per kilogram (kg) of body weight and about 400 mg per kg of body weight is
0 sl- --'le P~,fe.ably from 15 mg to about 150 mg/kg of body weight is used.
Generally, the dosage in man is lower than for small warrn blooded .. _.. ~lc such as
mice. A dosage unit may co.-.~,.ise a single compound or mixtures thereofwith other
comrolln~s or other cancer inhihiting comroun~lC The dosage unit can also co...~-ise
uçntC ~ n~ , carriers and the like. The unit may be in solid or gel forrm such as
15 pills, tablets, c~ps~ and the like or in liquid form suitable for oral, rectal, topical,
intravenous je tion or parenle~al ~d .. -- l~alion or inje~lion into or around the
tumor.
D. DOSAGE DELIVERY FORMS
The anti-cancer co--.~,ùu,-ds are typically mixed with a pha-..-~c~ ly
20 acce~L~I~ carrier. This carrier can be a solid or liquid and the type is generally chosen
based on the type of a ' ~ation being used. The active agent can be
co~ eu ~d in the form of a tablet or c-q-rsnl~, as an q-gglomerated powder or in a
liquid form. F - . ' of suitable solid carriers include lactose, sucrose, gelatin and
agar. Cqrs~ or tablets can be easily formlllqted and can be made easy to swallow or
25 chew, other solid forms include granules, and bullc p.,~.l~ ,. Tablets may contain
su~table binders, l..l" i~ s, di~ nts, ~ {~ y aLi~3 agents, c ~Ic .ng agents, na~ul-ng,
agenh, flow-;n-lu~ g agents, and melting agents. F . ' - s of suitable liquid dosage
forms include S''~ ~tinne or s~ n~ C!nC in water, phall.~ ,c~ ~;C'qlly ?~ ~r ~le fats and
oils, qlc4hnl~ or other organic solvents, inrl~lding esters, ~ ~.---I-'on~, syrups or eli~cirs,
30 s~l~rf ~ :on~, s~!-ltinnc and/or ,-~y~n- Ol-~ r~o~ ted from non-effervescent granules
and effervescent plepa-alions ,~on~ led from effervescent g.a.,ules. Such liquiddosage forms may contqin, for; . Ir, suitable solvents, ples~ ti.~es, c.--ulsir~l..g
agents, ;,..,~ 3 agents, ~ ents, ~ _ten~.:" th- ' ~, and melting agents. Oral
dosage forms optionally contain flavorants and coloring agents. P~e.ll~.al and
3s intravenous forrns would also include rninerals and other materials to make them
crJ",p~ til ' e with the type of j~ or delivery system chosen.
Specific ~ . ' ~ ~ of ph~ .- .~cc~l ;c~l ~q- ~cept~ carriers and .. . - that
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may be used to formulate oral dosage forrns of the present invention are des~,-;l,ed in
US. Pat. No. 3,903,297 to Robert, issued Sept. 2, 1975. Techniques and compositions
for making dosage forms useful in the present invention are des-,libed in the following
..,~,e.~ces: 7 Modern Pharm~ e~lti~s Chapters 9 and 10 (Banker & Rhodes, Editors,
s 1979); Lie~c.",dn et al., Pharm~seutiç~l Dosage Forrns: Tablets (1981); and Ansel,
Introduction to Pl,a,...~entic~sl Dosa~e Forrns 2nd Edition (1976).
E. MET~OD OF TREATMENT
The method of l,~ ..v ~ can be any suitable method which is effective in the
of the particular cancer type or virus that is being treated. Tr .. ~ may be
o oral, rectal, topical, pa, ~ le. ~l or intravenous ~ n alion or by injection into the
tumor and the like. The method of applying an effective amount also varies depPn~iing
on the tumor being treated. It is believed that parenteral t,_~ .l by intravenous,
~ b~ rous, or intr~m~c~ r applir~tiQn ofthe lH-1,2,4-tri~ole cG",po~"ds,
fo~ teJ with an app,ol"iale carrier, a~dition~l cancer inhilJiting co~ o~nd or
lS CO~ O~ c or diluent to fi~cilit~te applic~tion will be the ~ref.,.ll,d method of
t~ u)~. the co~.l.u~ c to warm blooded ~nin slc
The method of ll~ling viral ;..r~;l;O!.c may also be by oral, rectal, topical,
p&e.lt~.~l or intravenous ~ alion.
In vitro Data
The following e - ~ k ~ are illustrative and are not meant to be limiting to theinvention.
Colon~ Breast and Lung Tumor Cells Test
The follo~,u~, cell culture tests were p_.ru.,l,ed to test the toxicity of the N-
F~h~ hon~ u~c eQ~ C on colon, breast and lung human tumor cells. The
2s viability ofthe cells werc tested by looking at M'rT (3-t4,5-du,.~,ll,ylllùazol-2-yl] -2,5-
di~ t~.t~ " m bl~ '1e)ri-hl~,l;OIl MTT assay is a well known ",~le of cell
viabil~.
The colon tumor cells (E~29 from American Type Culture Colle~ion (ATCC)
) and the breast cells (MX1 from cell lines from ATCC) were cultured in Eagle's
Miminsl ~.cc~ntis~l Mc-l:-~--- with 10% fetal bovine serum. The lung tumor cells (A549
from ATCC cell lines) were cultured in Ham's F12 m~il-m with 10% fetal bovine
serwn
The tumor cells were p~c~ d and seeded into culture flasks at the desired cell
;es The culture mP~ m was ~le~ ed and the cell sheets were washed twice
with phospb~te buffered saline (PBS). The cells were tT~y~ d and lliLulale;l prior
to seeding the flasks. Unless otherwise i~ d the cultures were i~ b ~ at 37 +
1~ C in a I ' '' ~ ss~,h ~; of 5+ 1% carbon dioxide in air. The cultures were
=-- .
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inc~lb~d until they were 50-80% confluent.
The cells were subc~ ed when the flasks were stlbco~flllPnt The .,.P.li.....
was as~,i,alcd from the flasks and the cell sheets rinsed twice with PBS. Next, the
Trypsin Solution was added to each flask to cover the cell sheet. The Trypsin Solution
5 was removed after 30-60 seCon~c and the flasks were inC~b~tpd at room te-"pe,al,lre
for two to six .~ .. s When 90% of the cells became di~lodgP~ growth mP~i~lm wasadded. The cells were removed by trituration and ll~ sr~.led to a sterile centrifuge
tube. The co~c~ alion of cells in the sllcpPn ;on was deh~"~med~ and an approp, ;àle
dilution was made to obtain a density of 5000 cells/ml. The cells were sul~ d into
the d~ A~d wells ofthe 96-well bioassay plates (200 ,~ tol~ r cell ~ or~ per
well). PBS was added to all the ~ ini~g wells to ..~ ;n humidity. The plates
were then inrllb~t-pd overnight before test article ~ " ~
Each dose of test article was tested by ll~ia~ qud-ll .,p!ic ~e wells of cultures
with lO0 microliter of each ~illltiorl Those wells d~ ~i as solvent co~lllols
15 received an ~ ' 100 m ( ol;l~r of ...~ nol control; negative colltlols wells
received an ?~ 00 ~ 'ù~ oftr~ medillm PBS was added to the
re ~ ~ wells not treated with test article or ",~ The plates were then
,Jl,s~l for appro~ t~ly S days.
At the end ofthe 5 day ;...-~lb_l;on, each dose group was ~
20 Ill..,rosco~ lly to assess toxicity. A 0. 5 mg/ml dilution of Mrr was made in... .t ,..r.1i~l..~, and the dilution was filtered through a 0,.45 ~ u..._t~r filter to
remove l~n~l;c~lved crystals. The .,.~.1;.~.,, was deç-~t~d from the wells ofthe bioassy
plates. ~ ' Iy ~ r~L,r, 2000 1l~,_l ol;ler of the filtered MTT s Dll~tinn was added
to aU test wells except for the two u~lt~,aled blank test wdls. The two blank wells
2s ro~;,_d 200 microliters of 1, "1-n~ .-.e l;----- The plates were r~lulllçd to the
~ ' ~,~ r for a}~ hours. A~er in~b~tion the MrT co.-l-;n ~g ...~ was
dec~ed. Exces~ ",~ ~-,. was added to each well and the plates were shaken at room
t~np~~ue for about 2 hours.
The abs ~, ~ance at 550 nm (OD550) of each well was u.eas~rcd with a
30 M~!e ' - Devices ~Menlo Park, CA) VMax plate reader.
The mean ODsso of the solvent control wells and that of each test article
ltion, and that of each of the blank wells and the positive control were ç~ ted
The mean ODsso of the blank wells was sul~ from the mean of the solvent
control wells, and test article wells, lespc_~i~ely to give the co-l~ ,ondil.g mean
3s OD550
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% of Control = co.-~,cLed mean OD~50 of Test Article Dilution X 100
cO~ ed mean of OD550 of Solvent Control
Dose f_S~JOI~G curves were pn,p&ed as semi-log plots with % of control on the
o.di. ale (linear) and the test article collr~ ion on the ~bsc:c~ (log~ ' ~). The
s ECso was i~Lel~olaled from the plots for each test article.
For the test articles ~ ..;.. c~.ed in ...~ It~nol~ sepalale responses were
prepan,d to correct for the ~- ~el 1-~ nol data.
Adl;alll~cill was used as a positive control. In all cases, it was more toxic than
any of the test materials by one or two logs. Ad.i~.-~i n is one of the more potent
0 agents in current use and one with ~ r~ side effects. The peak plasrna
conçP~n~lion of other, quite effective cl~ OIll~,.ap~,utic agents may be 10 to 50 times
higher than that of Ad~ -. The EC-50 is the conr,~ alion at which one half the
cells are killed.
Table 1
Test MaterialEC-50 Result Cppml)
HT29 MX1 A549
Ad~;alll~;ill 0.00639 0.00078 0.00373
Flo~ le û.û33 i û.û284 0. i i3
These cAI.~,,illl~,n~s show that these compositions are effective in killing tumor
cells without !~ gJ.;~ y ~ healthy cells. They are safer than ad.;~.. ~,i-
